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发明公开
CN107841490A 双功能亚甲基四氢叶酸脱氢酶/环化酶及其多克隆抗体 有权

双功能亚甲基四氢叶酸脱氢酶/环化酶及其多克隆抗体
摘要：
本发明涉及分子生物学领域，尤其涉及一种双功能亚甲基四氢叶酸脱氢酶/环化酶及其多克隆抗体。双功能亚甲基四氢叶酸脱氢酶/环化酶的多克隆抗体的制备方法，包括步骤：1)以氨基酸序列如SEQ ID No.1所示的双功能亚甲基四氢叶酸脱氢酶/环化酶为抗原，免疫小鼠制备抗血清；2)纯化抗血清得到双功能亚甲基四氢叶酸脱氢酶/环化酶的多克隆抗体。本发明利用PCR技术在喜冷杆菌属Cryobacterium中扩增得到双功能亚甲基四氢叶酸脱氢酶/环化酶基因，并通过基因工程技术得到Ecoli BL21-pET21a-BMTHC重组菌株，诱导表达重组菌株得到双功能亚甲基四氢叶酸脱氢酶/环化酶蛋白，该蛋白具有较好的低温耐受，而且以双功能亚甲基四氢叶酸脱氢酶/环化酶为抗原成功得到双功能亚甲基四氢叶酸脱氢酶/环化酶的多克隆抗体。
摘要（英）：

The invention relates to the field of molecular biology, in particular to difunctional methylenetetrahydrofolate dehydrogenase/cyclase and a polyclonal antibody thereof. A preparation method of the polyclonal antibody of the difunctional methylenetetrahydrofolate dehydrogenase/cyclase comprises the steps of (1) adopting the difunctional methylenetetrahydrofolate dehydrogenase/cyclase with an aminoacid sequence as shown in SEQ ID NO.1 as an antigen, and immunizing mice for preparing an antiserum; (2) purifying the antiserum to obtain the polyclonal antibody of the difunctional methylenetetrahydrofolate dehydrogenase/cyclase. According to the difunctional methylenetetrahydrofolate dehydrogenase/cyclase and the polyclonal antibody thereof provided by the invention, a PCR (Polymerase Chain Reaction) technology is utilized for amplifying in cryobacterium to obtain a difunctional methylenetetrahydrofolate dehydrogenase/cyclase gene, and an Ecoli BL21-pET21a-BMTHC recombination strain is obtained through a gene engineering technique and is inducibly expressed to obtain a difunctional methylenetetrahydrofolate dehydrogenase/cyclase protein. The protein has better low temperature endurance, and the polyclonal antibody of the difunctional methylenetetrahydrofolate dehydrogenase/cyclase is successfully obtained by adopting the difunctional methylenetetrahydrofolate dehydrogenase/cyclaseas the antigen.

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